Methods of reducing Ascogregarina chagasi parasitaemia in laboratory colonies of Lutzomyia longipalpis.

Ascogregarina chagasi is an aseptate gregarine parasite found naturally in populations of Lutzomyia longipalpis. Under intensive rearing conditions in the laboratory, the parasite is known to reduce longevity and egg production, therefore, it is thought to be a major contributing factor to the phenomenon of colony crashes. A study is described to test egg cleansing techniques, with a view to controlling this parasite in laboratory colonies. Methods used include washing eggs with formol, benzalkonium chloride and irradiating with ultraviolet light. It was concluded that cleansing with 0.1% formol solution was the most efficient method of controlling A. chagasi.

Successful immunization of humans with irradiated malaria sporozoites: humoral and cellular responses of the protected individuals.

Two groups of volunteers were vaccinated by repeated exposure to the bites of Plasmodium falciparum-infected, x-irradiated mosquitoes in order to characterize the humoral and cellular immune responses of sporozoite-immunized, protected individuals. One of the two volunteers in the first immunization trial, when challenged by the bite of P. falciparum-infected mosquitoes, developed an infection only after a prolonged prepatent period. A second group of three volunteers who were exposed more frequently to larger numbers of infected mosquitoes irradiated with a lower x-ray dose was completely protected against sporozoite challenge. These individuals and the volunteer with delayed infection had high levels of antibodies to sporozoites and to the repeat region of the circumsporozoite (CS) protein. The CS-specific cellular immune responses of these volunteers were also stimulated by sporozoite immunization, as determined by proliferation of peripheral blood mononuclear cells (PBMC) and mitogen or antigen-expanded PBMC, in response to in vitro challenge with a recombinant P. falciparum CS protein. Based upon the assays used in this study, it is not possible to reach conclusions regarding specific immunologic responses and protection from sporozoite challenge.

Immunization against bovine tropical theileriosis, using 60Co-irradiated infective particles of Theileria annulata (Dschunkowsky and Luhs 1904) derived from ticks.

Portions of ground supernate material from Theileria annulata-infected and prefed adults of the Hyalomma anatolicum anatolicum were irridiated by 60Co source at 0, 3, 5, 6, 7, or 9 krads. These portions were injected subcutaneously into different groups of cross-bred calves. The calves injected with 0-, 3-, or 5-krad irradiated tick supernate had severe reactions (rectal temperature 39.8 to 41.6 C, two- to fourfold enlargement of prescapular lymph gland, mean peak parasitemia 27.3% to 32.5% infected erythrocytes, decreased PCV 41.7% to 60.7%, and hemoglobin concentration 37.5% to 50.8%), and many died, whereas those injected with 6- or 7-krad irradiated tick supernate had minimal reactions (rectal temperature 39.2 to 39.8 C, slight enlargement of prescapular lymph gland, parasitemia less than 1%, decrease packed cell volume 15.1%, and hemoglobin concentration 20.1%), with no mortality. All surviving calves injected with nonirradiated (0-) or 3-, 5-, 6-, or 7-krad irradiated tick supernate developed comparable indirect fluorescent antibody titers and were resistant to challenge exposure with infective material from 30 ticks. The calves injected with 9-krad irradiated tick supernate showed no reactions and did not develop indirect fluorescent antibody titers; they were also susceptible to challenge exposure with infective tick material.

Immunization of man against falciparum and vivax malaria by use of attenuated sporozoites.

With strict adherence to ethical guidelines, a volunteer was immunized against sporozoites of Plasmodium falciparum and P. vivax, the antigen consisting of attenuated sporozoites of each species inoculated through bites of mosquitoes X-irradiated at a minimum dosage of 15,000 rads. On one occasion this dosage did not render all P. vivax sporozoites noninfective. Species specificity of antigen and antibody was demonstrated, but within each species a wide geographical diversity of strains proved interchangeably antigenic and susceptible to the antibody. Once immunized, the volunteer was protected for not more than 3 months and 6 months, respectively, from infective P. falciparum and P. vivax sporozoites, the duration of protection being reflected by a positive species-specific circumsporozoite reaction. Studies in this volunteer, and in two others immunized with P. falciparum sporozoites, did not reveal any increase in serum levels of immunoglobulins G and M.